Cell Signaling Technology

Product Pathways - Tyrosine Kinase/ Adaptors

Phospho-Dab1 (Tyr220) Antibody #3327

Applications Reactivity MW (kDa) Source
W H (M) (R) 80, 110 GFP-Dab1 fusion. Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Dab1 (Tyr220) Antibody detects transfected levels of Dab1 protein only when phosphorylated at tyrosine 220.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to residues surrounding Tyr220 of human Dab1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of cell lysates from COS cells co-transfected with GFP-Dab1 and Fyn expression constructs, untreated and treated with calf intestinal phosphatase (CIP), using Phospho-Dab1 (Tyr220) Antibody.

Background

The Reelin signaling pathway plays a critical role in neuronal development. Reelin is a secreted glycoprotein that binds to the lipoprotein receptors VLDLR and ApoER2 or alpha3beta1 integrin on the surface of neurons (1,2). Activation of these receptors induces tyrosine phosphorylation of Disabled 1 (Dab1), an intracellular adaptor. It is generally believed that tyrosine phosphorylation of Dab1 by Src family tyrosine kinases is the most critical downstream event in Reelin signaling. The phosphotyrosine-binding (PTB) domain within its amino terminus enables Dab1 to recognize and bind to a conserved sequence motif within the cytoplasmic tail of the receptors. In addition, the PTB contains a Pleckstrin Homology-like subdomain that binds to phosphoinositides. The phosphoinositide-binding region within the Dab1 PTB domain is required for membrane localization and basal tyrosine phosphorylation of Dab1 independent of VLDLR and ApoER2 (3). It has been demonstrated that Src, CrkII, CrkL and Dock1 associate with tyrosine-phosphorylated Dab. The CrkII-Dab1 interaction requires tyrosine phosphorylation of Dab1 at residues 220 or 232 (4).

  1. Huang, Y. et al. (2005) Biochem. Biophys. Res. Commun. 331, 1460-1468.
  2. Luque, J.M. (2004) Brain Res. Dev. Brain Res. 152, 269-271.
  3. Morimura, T. et al. (2005) J. Biol. Chem. 280, 16901-16908.
  4. Chen, K. et al. (2004) J. Cell. Sci. 117, 4527-4536.

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